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  • LongAmp™ Taq 2X Master Mix

    Description

    The LongAmp® Taq 2X Master Mix combines high quality NEB recombinant Taq DNA Polymerase with a trace amount of Deep VentR DNA Polymerase. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). This ready-to-use mix offers two fold higher fidelity than Taq DNA Polymerase alone. A wide range of PCR products can be generated, up to 30 kb from lambda or human genomic DNA.

    Amplification of specific sequences from human genomic DNA using LongAmp Taq DNA Polymerase. Amplicon sizes are indicated below the gel. Marker M is the NEB 1 kb DNA Ladder (NEB #N3232 ).

    Product Source

    An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep Vent DNA Polymerase gene from Pyrococcus species GB-D.

    Advantages and Features

    Applications

    • Long Range PCR
    • High Throughout PCR
    • Colony PCR

    Properties and Usage

    Storage Temperature

    -20°C

    Buffer Composition

    60 mM Tris-SO4
    20 mM (NH4)2SO4
    2 mM MgSO4
    0.3 mM dNTPs
    3% Glycerol
    0.06% IGEPAL® CA-630
    0.05% Tween® 20
    125 units/ml LongAmp Taq DNA Polymerase
    pH 9@25°C

    Heat Inactivation

    No

    Unit Assay Conditions

    1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.

    Reaction Conditions: 
    1X LongAmp Taq Master Mix, DNA template and primers in a total reaction volume of 25–50 µl.

    Unit Definition:
    One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 75°C.

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • PCR Amplification (DNA Polymerase):

      The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.

    Notes

    1. LongAmp Taq 2X Master Mix is stable for fifteen freeze-thaw cycles when stored at -20°C.
    2. LongAmp Taq 2X Master Mix is also stable for three months at 4°C, so for frequent use, an aliquot may be kept at 4°C.

    References

    1. Barnes, W. M. (1994). Proc. Natl. Acad. Sci. USA. 91, 2216-2220.
    2. Saiki R. K. et al. (1985). Science. 230, 1350-1354.
    3. Powell, L. M. et al. (1987). Cell. 50, 831-840.
    4. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques. 15, 372-374.
    5. Sarkar, G., Kapelner, S. and Sommer, S. S. (1990). Nucleic Acids Res. 18, 7465.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. What is the fidelity of the LongAmp™ Taq DNA Polymerase compared to Taq DNA Polymerase?
    2. What type of DNA ends result from a primer extension reaction or a PCR using LongAmp™ Taq DNA Polymerase?
    3. What is the extension rate when using LongAmp™ Taq DNA Polymerase?
    4. Can the extension step be carried out at 72°C when using LongAmp™ Taq DNA Polymerase?
    5. Can LongAmp™ Taq DNA Polymerase be used to amplify GC-rich amplicons?
    1. Protocol for LongAmp™ Taq 2X Master Mix

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools

    Using an extension temperature of 65°C is recommended in LongAmp Taq reactions.